Recovery of viable cells from rabbit skin biopsies after storage at −20°C for up to 10 days

Abstract

Frozen animal tissues are thought to be appropriate for use as a donor for somatic cell nuclear transfer. This makes the freezing for long term storage a valuable tool for breeders needing to protect an animal population that is endangered by sanitary problems or for cryobanking of genetic resources. We report the successful cryopreservation of explants of skin derived from small biopsies from rabbit ear biopsies by using a protocol that can be easily performed by usual breeders, which are not equipped with cooling devices. By optimizing the procedure, we show that small biopsies can be kept at -20°C in a physiological solution containing 10% DMSO for up to 20 days before being deeply frozen in liquid nitrogen for long-term storage. After 10 days of storage at -20°C, the rate of viability of biopsies was similar to the control one (86 and 82% respectively). After 20 days of storage at -20°C, the rate of viability was dramatically lowered (39%), but it still allows to recover a significant population of viable cells from the preserved sample. Being appropriate to places lacking specific device, such a very simple technique may contribute to facilitate genome banking policies dedicated to the management of genetic resources in wild and domestic animals.

Combination of ethylene glycol with sucrose increases survival rate after vitrification of somatic tissue of collared peccaries (Pecari tajacu Linnaeus, 1758)

“…In general, the cryopreservation is a strategy for genetic conservation of animals, domestic or wild (Benkeddache et al 2012) and vitrification procedures are superior to conventional cryopreservation by freezing (Brockbank et al 2010). In the vitrification, the solution is rapidly cooled and transformed into a glassy, vitrified state, not by ice crystallization, but because of extreme elevation in viscosity during cooling (Amorim et al 2011).…”